This study was done on a total of 92 mastitic milk samples (50 clinical, 42 sub-clinical) and 40 hand swabs from contact humans  were collected from different dairy farms at  Gharbia governorate.  The collected samples were examined bacteriologically to isolate mastitis pathogens (Staph. aureus, Strept. agalactiae, S. dysgalactiae and Strept. uberis). From clinical mastitic samples, six isolates were  S. aureus ( 12%) and one isolate  was (2%) S. dysgalactiae.  Among sub clinical mastitic milk samples two isolates were S. aureus (4.6%) and one  isolate (2.3%) S. agalactiae. While S. uberis were not detected.  From contact human hand swabs both S. aureus and Streptococcus species were s were highly sensitive to Antibiotic sensitivity test revealed that all bacterial isolatenot detected. gentamicin respectively, while all isolates  andenerofloxacin, ciprofloxacin, sulpha |trimethoprim were resistant to penicillin followed by amoxicillin/Clavulanic acid. Two isolates of S. aureus were screened for detection of enterotoxin genes (Sea, Seb, Sec, Sed and See) by multiplex PCR. Only Sed gene was detected in one isolate.  Cfb gene (CAMP factor) and hyl (hyluronidase) gene were detected in S. agalactiae. mig (surface-expressed mig protein) gene was detected in S. dysgalactiae